Nato3 plays an integral role in dorsoventral patterning of the spinal cord by segregating floor plate/p3 fates via Nkx2.2 suppression and Foxa2 maintenance.

During embryogenesis, the dorsal roof plate and the ventral floor plate (FP) act as organizing centers to pattern the developing neural tube. Organizer-secreted morphogens provide signals that are interpreted via the graded expression of transcription factors. These factors establish a combinatorial code, which subsequently determines the fate of neuronal progenitors along the dorsoventral axis. To further separate the fates and promote distinct identities of the neural progenitors, mutual repression takes place among transcription factors expressed in progenitors situated along the dorsoventral axis. The molecular mechanisms acting in the developing spinal cord and underlying the segregation of the progenitor pool containing cells with a mixed FP/p3 fate into separate FP cells and V3 neurons are not fully understood. Using in vivo ectopic expression in chick, we found that Nato3 induces ectopic Foxa2-positive cells and indirectly downregulates Nkx2.2 expression. To examine the role of Nato3 in the FP, Foxa2-Nato3 signaling was blocked in Nato3 null mice and to a greater extent in Nato3 null/Foxa2 heterozygous bigenic mutants. Complementary to the findings obtained by gain of function in chick, the loss of function in mouse indicated that the segregation of the FP/p3 population into its derivatives was interrupted. Together, the data suggest that Nato3 is a novel determinant factor regulating the segregation of the FP and p3 identities, which is an essential step for establishing a definitive FP fate in the embryonic spinal cord.

Floor plate descendants in the ependyma of the adult mouse Central Nervous System.

During embryonic development of the Central Nervous System (CNS), the expression of the bHLH transcription factor Nato3 (Ferd3l) is unique and restricted to the floor plate of the neural tube. In mice lacking Nato3 the floor plate cells of the spinal cord do not fully mature, whereas in the midbrain floor plate, progenitors lose some neurogenic activity, giving rise to a reduced population of dopaminergic neurons. Since the floor plate is considered to be disintegrated at the time of birth, Nato3 expression was never tested postnatally and in adult mice. Here, we utilized a Nato3 knockout mouse model in which a LacZ reporter precisely replaced the coding region under the endogenous regulatory elements, so that its expression recapitulates the spatiotemporal pattern of Nato3 expression. Nato3 was found to be expressed in the CNS throughout life in a highly restricted manner along the medial cavities: in subpopulations of cells in the IIIrd ventricle, the cerebral aqueduct, the IVth ventricle, the central canal of the spinal cord, and the subcommissural organ, a gland located in the midbrain. A few unifying themes are shared among all Nato3-positive cells: all are positioned in the midline, are of an ependymal type, and contact the cerebrospinal fluid (CSF) similarly to the embryonic position of the floor plate bordering the lumen of the neural tube. Taken together, Nato3 defines an unrecognized subpopulation of medial cells positioned at only one side of circular ependymal structures, and it may affect their regulatory activities and neuronal stem cell function.